Identification of host-targeted small molecules that restrict intracellular Mycobacterium tuberculosis growth.
Identification of host-targeted small molecules that restrict intracellular Mycobacterium tuberculosis growth.
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DOI：
10.1371/journal.ppat.1003946
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发表时间：
2014-02
期刊：
影响因子：
6.7
通讯作者：
Hung DT
中科院分区：
文献类型：
--
作者：
Stanley SA;Barczak AK;Silvis MR;Luo SS;Sogi K;Vokes M;Bray MA;Carpenter AE;Moore CB;Siddiqi N;Rubin EJ;Hung DT
结核分枝杆菌仍然是对全球健康的重大威胁。巨噬细胞是M.结核感染，并且尽管细菌能够在某些条件下在细胞内复制，但也清楚的是巨噬细胞能够杀死M.肺结核如果适当激活。感染的结果至少部分取决于巨噬细胞内宿主与病原体的相互作用;然而，我们缺乏对哪些宿主途径对细菌生存和复制至关重要的完整了解。为了增加我们对细胞内感染所涉及的分子过程的理解，我们使用高含量显微镜测定进行了化学筛选，以通过靶向宿主功能和途径来鉴定限制分枝杆菌在巨噬细胞中生长的小分子。鉴定的宿主靶向抑制剂仅在巨噬细胞感染的情况下限制细菌生长，并且主要分为五类：G蛋白偶联受体调节剂、离子通道抑制剂、膜转运蛋白、抗炎剂和激酶调节剂。我们发现，氟西汀（一种选择性5-羟色胺再摄取抑制剂）可增强促炎细胞因子TNF-α的分泌，并诱导受感染巨噬细胞的自噬，而吉非替尼（一种表皮生长因子受体（EGFR）抑制剂）也可激活自噬并限制生长。我们证明，在感染过程中，通过EGFR的信号激活p38 MAPK信号通路，防止巨噬细胞有效地响应感染。在体内感染期间使用吉非替尼抑制该途径可减少M.在受感染的小鼠的肺中发现结核。我们的研究结果支持这样的概念，即使用细胞内模型筛选抑制剂导致在体内感染期间识别用于探测途径的工具化合物，并且还可能导致识别通过调节宿主途径起作用的新的抗结核药物。鉴于我们鉴定的一些化合物用于其他治疗适应症的现有经验，值得对这些化合物进行进一步的临床导向研究。细菌病原体结核分枝杆菌（Mycobacterium tuberculosis）的感染导致结核病（TB），其在世界范围内造成显著的发病率和死亡率。大约有20亿人感染了M。结核病，每年有近150万人死于结核病。随着耐药性的增加和新的候选药物很少，我们无法有效地治疗所有感染的个体，需要更深入地了解宿主-病原体界面，以促进新的治疗方法。此外，目前的抗结核治疗方案需要数月的严格依从才能清除感染;靶向宿主免疫功能可以在有效治疗耐药菌株的同时，在减少治疗时间和复杂性方面发挥战略作用。在这里，我们使用基于显微镜的筛选来鉴定靶向宿主途径并抑制M生长的分子。巨噬细胞中的结核病。我们确定了几种以前与结核病无关的宿主途径。所鉴定的抑制剂通过阻断M.结核病的毒力，或通过激活针对细胞内细菌的免疫反应。临床上用于治疗抑郁症的氟西汀诱导自噬并增强TNF-α的产生。类似地，临床上用于治疗癌症的吉非替尼抑制M.结核菌在巨噬细胞中生长。重要的是，吉非替尼治疗减少了M.感染结核病的小鼠。
Mycobacterium tuberculosis remains a significant threat to global health. Macrophages are the host cell for M. tuberculosis infection, and although bacteria are able to replicate intracellularly under certain conditions, it is also clear that macrophages are capable of killing M. tuberculosis if appropriately activated. The outcome of infection is determined at least in part by the host-pathogen interaction within the macrophage; however, we lack a complete understanding of which host pathways are critical for bacterial survival and replication. To add to our understanding of the molecular processes involved in intracellular infection, we performed a chemical screen using a high-content microscopic assay to identify small molecules that restrict mycobacterial growth in macrophages by targeting host functions and pathways. The identified host-targeted inhibitors restrict bacterial growth exclusively in the context of macrophage infection and predominantly fall into five categories: G-protein coupled receptor modulators, ion channel inhibitors, membrane transport proteins, anti-inflammatories, and kinase modulators. We found that fluoxetine, a selective serotonin reuptake inhibitor, enhances secretion of pro-inflammatory cytokine TNF-α and induces autophagy in infected macrophages, and gefitinib, an inhibitor of the Epidermal Growth Factor Receptor (EGFR), also activates autophagy and restricts growth. We demonstrate that during infection signaling through EGFR activates a p38 MAPK signaling pathway that prevents macrophages from effectively responding to infection. Inhibition of this pathway using gefitinib during in vivo infection reduces growth of M. tuberculosis in the lungs of infected mice. Our results support the concept that screening for inhibitors using intracellular models results in the identification of tool compounds for probing pathways during in vivo infection and may also result in the identification of new anti-tuberculosis agents that work by modulating host pathways. Given the existing experience with some of our identified compounds for other therapeutic indications, further clinically-directed study of these compounds is merited. Infection with the bacterial pathogen Mycobacterium tuberculosis causes the disease tuberculosis (TB) that imposes significant worldwide morbidity and mortality. Approximately 2 billion people are infected with M. tuberculosis, and almost 1.5 million people die annually from TB. With increasing drug resistance and few novel drug candidates, our inability to effectively treat all infected individuals necessitates a deeper understanding of the host-pathogen interface to facilitate new approaches to treatment. In addition, the current anti-tuberculosis regimen requires months of strict compliance to clear infection; targeting host immune function could play a strategic role in reducing the duration and complexity of treatment while effectively treating drug-resistant strains. Here we use a microscopy-based screen to identify molecules that target host pathways and inhibit the growth of M. tuberculosis in macrophages. We identified several host pathways not previously implicated in tuberculosis. The identified inhibitors prevent growth either by blocking host pathways exploited by M. tuberculosis for virulence, or by activating immune responses that target intracellular bacteria. Fluoxetine, used clinically for treating depression, induces autophagy and enhances production of TNF-α. Similarly, gefitinib, used clinically for treating cancer, inhibits M. tuberculosis growth in macrophages. Importantly, gefitinib treatment reduces bacterial replication in the lungs of M. tuberculosis-infected mice.
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DOI：
10.1084/jem.20080767
发表时间：
2008-11-24
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Chen M;Divangahi M;Gan H;Shin DS;Hong S;Lee DM;Serhan CN;Behar SM;Remold HG
通讯作者：
Remold HG
影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
DOI：
10.4049/jimmunol.1201538
发表时间：
2012-10-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Bruns H;Stegelmann F;Fabri M;Döhner K;van Zandbergen G;Wagner M;Skinner M;Modlin RL;Stenger S
通讯作者：
Stenger S
影响因子：
4.4
作者：
Cloëz-Tayarani, I;Petit-Bertron, AF;Cavaillon, JM
通讯作者：
Cavaillon, JM
影响因子：
64.5
作者：
Kumar, Dhiraj;Nath, Lekha;Rao, Kanury V. S.
通讯作者：
Rao, Kanury V. S.